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Factor D is the smallest complement protein with the lowest concentration and only one natural substrate with a high degree of selective function. It has emerged as an attractive target for effective and specific inhibition of alternative pathways. These features highlight the potential of factor D as an alternative pathway-mediated therapeutic target for disease. Creative Biolabs summarizes the CFD functional assay protocol below for your reference.
CFD Functional Assay Protocol
Factor D is a component of the complement replacement pathway and is famous for its role in the humoral inhibition of infection factors. It is a serine protease that stimulates glucose transport and inhibits fat decomposition.
Fig.1 Flow chart of CFD functional assay protocol. (Creative Biolabs)
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Fig.2 Supplementation of CFD-deficient serum with purified CFD accelerates complement activation in a dose-dependent manner.1
Correct measurement of CFD activity remains essential for both the assessment of therapeutic inhibitors and the identification of complement-mediated diseases. Current technological developments focus on utilizing fluorogenic substrates to measure enzymatic cleavage in real-time, which facilitates high-throughput screening while improving sensitivity. The optimization of substrate specificity and reaction kinetics alongside inhibitor potency takes place with precision to reduce background noise in measurements. The use of purified CFD combined with physiological buffers like PBS at pH 7.4 and maintained at 37°C within protocols produces experimental conditions that accurately mimic natural physiological activity. Validating results through parallel ELISA and functional hemolytic assays confirms target engagement and specificity while improving data reliability.
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With our team of innovative technologies and experts, Creative Biolabs is supporting the scientific community in achieving its goals, advancing its knowledge of complement functions, and making discoveries. Please contact us to discuss the research project you are interested in. Our team will introduce our complement testing services and product information in detail, to provide you with reasonable suggestions that meet your requirements.
Reference
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Langereis, Jeroen D., et al. "Complement factor D haplodeficiency is associated with a reduced complement activation speed and diminished bacterial killing." Clinical & Translational Immunology 10.4 (2021): e1256. Distributed under an Open Access license CC BY 4.0, without modification.
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